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2009 Fiscal Year Final Research Report

Elucidation of Cl--pumping mechanism and significance of trimer formation of halorhodopsin

Research Project

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Project/Area Number 19390009
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Physical pharmacy
Research InstitutionHokkaido University

Principal Investigator

KAMO Naoki  Hokkaido University, 薬学部, 教授 (10001976)

Co-Investigator(Kenkyū-buntansha) KIKUKAWA Takashi  北海道大学, 大学院・先端生命科学研究院, 教授 (20281842)
NARA Toshifumi  松山大学, 薬学部, 准教授 (30241350)
MIYAUCHI Seiji  松山大学, 薬学部, 教授 (30202352)
Project Period (FY) 2007 – 2009
Keywordsハロロドプシン / センソリーロドプシン / フォボロドプシン / レチナールタンパク / 高度好塩菌 / オーサイト / フォトサイクル / 閃光光分解
Research Abstract

Halorhodopsin (HR) is a light-driven Cl^- pump expressed in the membrane of Halobacteria. We succeeded in developing a new method : HR was expressed in Xenopus laevis Oocytes, and photo-induced membrane current was measured due to the Cl^- transport. This method is highly quantitative. We prepared various HR mutants using E. coli expression system. Careful investigation on both the photochemistry and transport activities of the mutants has been done. We found Arg123 that is an essentially important amino acid residue. During the transition from N to O intermediate, Cl is released to cytoplasm while Cl is uptaken from extracellular space during O to HR' transition. We showed that HR forms trimer even in the presence of detergents, and that the trimer is more robust than the monomer.

  • Research Products

    (39 results)

All 2010 2009 2008 2007 Other

All Journal Article (24 results) Presentation (13 results) Book (1 results) Remarks (1 results)

  • [Journal Article] The photochemical reaction cycle and photoinduced proton transfer of sensory rhodopsin II2010

    • Author(s)
      J. Tamogami, T. Kikukawa, Y. Ikeda, A. Takemura, M. Demura, N. Kamo
    • Journal Title

      Halobacterium salinarum. Biophys. J. 8

      Pages: 1353-1363

  • [Journal Article] Effect of molecular assembly on photocycle of reconstituted bacteriorhodopsin: Significant blue shift of the late photointermediate in the liquid crystalline phase.2009

    • Author(s)
      M. Sonoyama, T. Kikukawa, Y. Yokoyama, M. Demura, N. Kamo, S. Mitaku
    • Journal Title

      Chemistry Lett. 39

      Pages: 1134-1135

  • [Journal Article] Production of functional bacteriorhodopsin by an Escherichia coli cell-free protein synthesis system supplemented with steroid detergent and lipid2009

    • Author(s)
      K. Shimono, M. Goto, T. Kikukawa, S. Miyauchi, M. Shirouzu, N. Kamo, S. Yokoyama
    • Journal Title

      Protein Sci. 18

      Pages: 2160-2171

  • [Journal Article] A simple and sensitive method for detection of Bacillus anthracis by loop-mediated isothermal amplification2009

    • Author(s)
      Y. Kurosaki, T. Sakuma, A. Fukuma, Y. Fujinami, K. Kawamoto, N. Kamo, S-I. Makino, J. Yasuda
    • Journal Title

      J. Appl. Microbiol. 107

      Pages: 1947-1956

  • [Journal Article] Effect of chloride binding on the thermal trimer-monomer conversion of halorhodopsin in the solubilized system.2009

    • Author(s)
      T. Sasaki, T. Aizawa, M. Kamiya, T. Kikukawa, K. Kawano, N. Kamo, M. Demura
    • Journal Title

      Biochemistry 48

      Pages: 12089-12095

  • [Journal Article] Reaction dynamics of halorhodopsin studied by time-resolved diffusion2009

    • Author(s)
      K. Inoue, M. Kubo, M. Demura, N. Kamo, M. Terazima
    • Journal Title

      Biophys. J. 96

      Pages: 3724-3734

  • [Journal Article] Role of Arg123 in light-driven anion pump mechanism of pharaonis halorhodopsin.2009

    • Author(s)
      M. Kubo, T. Kikukawa, S. Miyauchi, A. Seki, M. Kamiya, T. Aizawa, K. Kawano, N. Kamo, M. Demura
    • Journal Title

      Photochem. Photobiol. 85

      Pages: 547-555

  • [Journal Article] A tin oxide transparent electrode provides the means for rapid time-resolved pH measurements: Application to photoinduced proton transfer of bacteriorhodopsin and proteorhodopsin2009

    • Author(s)
      J. Tamogami, T. Kikukawa, S. Miyauchi, E. Muneyuki, N. Kamo
    • Journal Title

      Photochem. Photobiol. 85

      Pages: 578-589

  • [Journal Article] Halorhodopsin from Natronomonas pharaonis forms a trimer even in the presence of a detergent, dodecyle-β-D-maltoside2009

    • Author(s)
      T. Sasaki, M. Kubo, T. Kikukawa, M. Kamiya, T. Aizawa, K. Kawano, N. Kamo, M. Demura
    • Journal Title

      Photochem. Photobiol. 85

      Pages: 130-136

  • [Journal Article] Proton release group of pharaonis phoborhodopsin revealed by ATR-FTIR spectroscopy.2009

    • Author(s)
      Y. Kitade, Y. Furutani, N. Kamo, H. Kandori
    • Journal Title

      Biochemistry 48

      Pages: 1595-1603

  • [Journal Article] The lifetimes of pharaonis phoborhodopsin signaling states depend on the rates of proton transfer: Effects of hydrostatic pressure and stopped flow experiments2008

    • Author(s)
      T. Kikukawa, C.K. Saha, S.P. Balashov, E.S. Imasheva, D. Zaslavsky, R.B. Gennis, T. Abe, N. Kamo
    • Journal Title

      Photochem. Photobiol. 84

      Pages: 880-888

  • [Journal Article] A long-lived M-like state of phoborhodopsin that mimics the active state2008

    • Author(s)
      Y. Sudo, T. Nishihori, M. Iwamoto, K. Shimono, C. Kojima, N. Kamo
    • Journal Title

      Biophys. J. 95

      Pages: 753-760

  • [Journal Article] Dynamics change of phoborhodopsin and transducer by activation: study using D75N mutant of the receptor by site-directed solid-state 13C NMR2008

    • Author(s)
      I. Kawamura, H. Yoshida, Y. Ikeda, S. Yamaguchi, S. Tuzi, H. Saito, N. Kamo, A. Naito
    • Journal Title

      Photochem. Photobiol. 84

      Pages: 921-930

  • [Journal Article] Protein-protein interaction of a pharaonis Halrhodopsin mutant forming a complex with pharaonis halobacterial transducer protein II detected by Fourier-transform infrared spectroscopy.2008

    • Author(s)
      Y. Furutani, M. Ito, Y. Sudo, N. Kamo, H. Kandori
    • Journal Title

      Photochem. Photobiol. 84

      Pages: 874-879

  • [Journal Article] Structural changes in the O-decay accelerated mutants of pharaonis phoborhodopsin.2008

    • Author(s)
      Y. Sudo, Y. Furutani, M. Iwamoto, N. Kamo, H. Kandori
    • Journal Title

      Biochemistry 47

      Pages: 2866-2874

  • [Journal Article] Anti-parallel membrane topology of a homo-dimeric multidrug transporter, EmrE.2007

    • Author(s)
      T. Nara, T. Kouyama, Y. Kurata, T. Kikukawa, S. Miyauchi, N. Kamo
    • Journal Title

      J. Biochem. 142

      Pages: 621-625

  • [Journal Article] Structural analysis of the phototactic transducer protein HtrII liker region from Natronomonas pharaonis.2007

    • Author(s)
      K. Hayashi, Y. Sudo, J. Jee, M. Mishima, H. Hara, N. Kamo, C. Kojima
    • Journal Title

      Biochemistry 46

      Pages: 14380-14390

  • [Journal Article] Anti-parallel membrane topology of two components of EbrAB, a multidrug transporter2007

    • Author(s)
      T. Kikukawa, S. Miyauchi, T. Araiso, N. Kamo, T. Nara
    • Journal Title

      Biochem. Biophys. Res. Commun. 358

      Pages: 1071-1075

  • [Journal Article] Dynamics of light-induced conformational changes of the phoborhodopsin/transducer complex formed in the n-dodecyl β-D-Maltoside micelle.2007

    • Author(s)
      Y. Taniguchi, T. Ikehara, N Kamo, H. Yamasaki, Y. Toyoshima
    • Journal Title

      Biochemistry 46

      Pages: 5349-5357

  • [Journal Article] Participation of the surface structure of pharaonis phoborhodopsin, ppR and its A149S and A149V mutants. Consisting of the C-terminal □-helix and E-F loop, in the complex-formation with the cognate transducer pHtrII, as revealed by site-directed 13C solidParticipati2007

    • Author(s)
      I. Kawamura, Y. Ikeda, Y. Sudo, M. Iwamoto, K. Shimono, S. Yamaguchi, S. Tuzi, H. Saito, N. Kamo, A. Naito
    • Journal Title

      Photochem. Photobiol. 83

      Pages: 339-345

  • [Journal Article] Tryptophan 171 in pharaonis phoborhodopsin (sensory rhodopsin II) interacts with the chromophore retinal and its substitution with alanine or threonine slowed down the decay of M-and O-intermediate.2007

    • Author(s)
      T. Iwasa, E. Abe, Y. Yakura, H. Yoshida, N. Kamo
    • Journal Title

      Photochem. Photobiol. 83

      Pages: 328-335

  • [Journal Article] Interaction of the halobacterial transducer to a halorhodopsin mutant engineered so as to bind the transducer: Cl-circulation within the extracellular channel.2007

    • Author(s)
      C. Hasegawa, T. Kikukawa, S. Miyauchi, A. Seki, Y. Sudo, M. Kubo, M. Demura, N. Kamo
    • Journal Title

      Photochem. Photobiol. 83

      Pages: 293-302

  • [Journal Article] Application of fluorescence resonance energy transfer (FRET) to investigation of light-induced conformational changes of the phoborhodopsin/transducer complex2007

    • Author(s)
      Y. Taniguchi, T. Ikehara, N. Kamo, Y. Watanabe, H. Yamasaki, Y. Toyoshima
    • Journal Title

      Photochem. Photobiol. 83

      Pages: 311-316

  • [Journal Article] Heterologous expression of pharaonis halorhodopsin in Xenopus laevis oocyte and electrophysiological characterization of its light-driven Cl-pump activity2007

    • Author(s)
      A. Seki, S. Miyauchi, S. Hayashi, T. Kikukawa, M. Kubo, M. Demura, V. Ganapathy, N. Kamo
    • Journal Title

      Biophys. J. 92

      Pages: 2559-2569

  • [Presentation] 固体NMRによる光受容タンパク質ppRとその変異体T204Aの局所構造変化の解析2009

    • Author(s)
      T.Hidaka, et al.
    • Organizer
      第47回日本生物物理学会
    • Place of Presentation
      アスティ徳島
    • Year and Date
      2009-10-30
  • [Presentation] 固体13C NMRによるppRの細胞質表面部位の相互作用変化の観測2009

    • Author(s)
      T.Kondo, et al.
    • Organizer
      第47回日本生物物理学会
    • Place of Presentation
      アスティ徳島
    • Year and Date
      2009-10-30
  • [Presentation] 大腸菌発現系を用いたsalinarumハロロドプシンの機能的再構成2009

    • Author(s)
      Y.Yamashita, et al.
    • Organizer
      第47回日本生物物理学会
    • Place of Presentation
      アスティ徳島
    • Year and Date
      2009-10-30
  • [Presentation] 共鳴ラマン分光法によるファラオニスフォボロドプシンN中間体の検出2009

    • Author(s)
      M.Unno, et al.
    • Organizer
      第47回日本生物物理学会
    • Place of Presentation
      アスティ徳島
    • Year and Date
      2009-10-30
  • [Presentation] Halobacterium salinarum由来のフォボロドプシン(センサリーロドプシンII)の光化学反応サイクルとプロトン移動2009

    • Author(s)
      J.Tamogami, et al.
    • Organizer
      第47回日本生物物理学会
    • Place of Presentation
      アスティ徳島
    • Year and Date
      2009-10-30
  • [Presentation] ファラオニスハロロドプシンのCl輸送機構におけるAsp252の役割2009

    • Author(s)
      T.Kikukawa, et al.
    • Organizer
      第47回日本生物物理学会
    • Place of Presentation
      アスティ徳島
    • Year and Date
      2009-10-30
  • [Presentation] Analysis of photo-induced proon uptake/release by bacteriorhodopsin and proteorhodopsin using a SnO2 transparent electrode2008

    • Author(s)
      J. Tamogami, T. Kikukawa, S. Miyauchi, N. Kamo
    • Organizer
      13th International Conference on Retinal Proteins
    • Place of Presentation
      Barcelona. Invited Lecture
    • Year and Date
      20080615-20080619
  • [Presentation] Trimer assembly of Natronomonas pharaonis halorhodopsin, NpHR in lipid bilayer and detergent2008

    • Author(s)
      T. Tsukamoto, T. Sasaki, T. Kikukawa, M. Kamiya, T. Aizawa, K. Kawano, N. Kamo, M. Demura
    • Organizer
      13th International Conference on Retinal Proteins.
    • Place of Presentation
      Barcelona
    • Year and Date
      20080615-20080619
  • [Presentation] Photoreaction of salinarum phoborhodopsin studied by low-temperataure spectroscopy.2008

    • Author(s)
      D. Gang, Y. Ohno, Y. Ikeda, N. Kamo, T. Iwasa
    • Organizer
      13th International Conference on Retinal Proteins
    • Place of Presentation
      Barcelona
    • Year and Date
      20080615-20080619
  • [Presentation] Important amino acid residues of functioning in Halorhodopsin from Natronomonas pharaonis, NpHR.2008

    • Author(s)
      N. Kamo, S. Miyauchi, T. Kikukawa, H. Shigemura, Y. Saito, S. Hayashi, M. Kubo, T. Kitagawa, M. Demura
    • Organizer
      13th International Conference on Retinal Proteins
    • Place of Presentation
      Barcelona
    • Year and Date
      20080615-20080619
  • [Presentation] Backbone conformations of retinal protein as studied by solid state NMR.2008

    • Author(s)
      I. Kawamura, J. Tanabe, T. Nisho, A. Wada, S. Tuzi, N. Kamo, A. Naito
    • Organizer
      13th International Conference on Retinal Proteins
    • Place of Presentation
      Barcelona
    • Year and Date
      20080615-20080619
  • [Presentation] Role of Arg123 in light-driven anion pump mechanisms of N. pharaonis halorhodopsin, NpHR.2008

    • Author(s)
      M. Kubo, T. Kikukawa, S. Miyauchi, M. Kamiya, T. Aizawa, K. Kawano, N. Kamo, M. Demura
    • Organizer
      13th International Conference on Retinal Proteins
    • Place of Presentation
      Barcelona, selected presentation
    • Year and Date
      20080615-20080619
  • [Presentation] Effect of molecular assembly on photointermediates of BR reconstituted into dimyristoyolphosphatidylcholine vesicles.2008

    • Author(s)
      M. Sonoyama, T. Kikukawa, Y. Yokoyama, M. Demura, N. Kamo, S. Mitaku
    • Organizer
      13th International Conference on Retinal Proteins
    • Place of Presentation
      Barcelona
    • Year and Date
      20080615-20080619
  • [Book] 薬学生のための生物物理化学入門

    • Author(s)
      加茂直樹、嶋林三郎
    • Publisher
      廣川書店
  • [Remarks]

    • URL

      http://ghp01.matsuyama-u.ac.jp/~yakugaku/teacher/#teacher09

URL: 

Published: 2011-06-18   Modified: 2016-04-21  

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